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目的探讨白细胞介素-34/集落刺激因子-1受体(IL-34/CSF-1R)在转化生长因子-β1(TGF-β1)诱导A549细胞发生上皮-间质转化(EMT)中的表达。方法体外培养A549细胞;CCK 8法检测不同浓度TGF-β1在不同时间点对A549细胞增殖的影响;选用5μg/L TGF-β1刺激A549细胞(0、12、24、48h),提取细胞蛋白和RNA,Western blotting法检测α-平滑肌肌动蛋白(α-SMA)、E-钙黏连蛋白(E-Cad)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、基质金属蛋白酶抑制剂-1(TIMP-1)蛋白的变化;Real-time PCR法检测IL-34、CSF-1R、MMP-2、MMP-9基因表达的变化。结果 TGF-β1对A549细胞的增殖无明显影响(P>0.05)。TGF-β1刺激A549细胞后,间质细胞标志性蛋白α-SMA表达升高,上皮细胞标志性蛋白E-Cad表达下降,纤维化相关的MMP-2蛋白表达升高,MMP-9蛋白表达先升高后下降,TIMP-1蛋白早期出现短暂性增高后,呈持续降低趋势(P<0.01)。IL-34mRNA表达逐渐升高,CSF-1R mRNA、MMP-2 mRNA、MMP-9 mRNA表达先升高后下降(P<0.01)。结论 TGF-β1诱导A549细胞的EMT过程中,存在IL-34/CSF-1R的动态表达。
Objective To investigate the expression of IL-34 / CSF-1R in epithelial-mesenchymal transition (EMT) induced by transforming growth factor-β1 (TGF-β1) in A549 cells . Methods A549 cells were cultured in vitro. The effects of different concentrations of TGF-β1 on the proliferation of A549 cells were detected by CCK-8 assay. A549 cells were treated with 5μg / L TGF-β1 (0,12,24,48h) RNA and Western blotting were used to detect the expressions of α-SMA, E-Cadherin, MMP-2, MMP-9 ), MMP-2 and MMP-9 were detected by real-time PCR. The changes of IL-34, CSF-1R, MMP- Results TGF-β1 had no significant effect on the proliferation of A549 cells (P> 0.05). After A549 cells were stimulated by TGF-β1, the expression of α-SMA was increased, the expression of E-Cad in epithelial cells was decreased, the expression of MMP-2 protein was increased, the expression of MMP-9 protein was After increasing and then decreasing, the TIMP-1 protein showed a transient increase in the early stage, which showed a decreasing trend (P <0.01). The expression of IL-34 mRNA increased gradually, while the expression of CSF-1R mRNA, MMP-2 mRNA and MMP-9 mRNA increased first and then decreased (P <0.01). Conclusion The dynamic expression of IL-34 / CSF-1R in A549 cells induced by TGF-β1 exists in EMT.